Effect of Small Molecule on ex vivo Expansion of Cord Blood Hematopoietic Stem Cells: A Concise Review
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MINI REVIEW
published: 09 April 2021
doi: 10.3389/fcell.2021.649115
Effect of Small Molecule on ex vivo
Expansion of Cord Blood
Hematopoietic Stem Cells: A
Concise Review
Soudeh Ghafouri-Fard 1 , Vahid Niazi 2 , Mohammad Taheri 3* and Abbas Basiri 3*
1
Department of Medical Genetics, Shahid Beheshti University of Medical Sciences, Tehran, Iran, 2 Department of Tissue
Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University
of Medical Sciences, Tehran, Iran, 3 Urology and Nephrology Research Center, Shahid Beheshti University of Medical
Sciences, Tehran, Iran
Hematopoietic stem cells (HSCs) are a group of cells being produced during
embryogenesis to preserve the blood system. They might also be differentiated to
non-hematopoietic cells, including neural, cardiac and myogenic cells. Therefore, they
have vast applications in the treatment of human disorders. Considering the restricted
Edited by:
Hiroki Shibata,
quantities of HSCs in the umbilical cord blood, inadequate mobilization of bone marrow
Kyushu University, Japan stem cells, and absence of ethnic dissimilarity, ex vivo expansion of these HSCs is
Reviewed by: an applicable method for obtaining adequate amounts of HSCs. Several molecules
Bradley Wayne Doble, such as NR-101, zVADfmk, zLLYfmk, Nicotinamide, Resveratrol, the Copper chelator
University of Manitoba, Canada
Wilairat Leeanansaksiri, TEPA, dmPGE2, Garcinol, and serotonin have been used in combination of cytokines
Suranaree University of Technology, to expand HSCs ex vivo. The most promising results have been obtained from cocktails
Thailand
that influence multipotency and self-renewal features from different pathways. In the
*Correspondence:
Mohammad Taheri
current manuscript, we provide a concise summary of the effects of diverse small
mohammad_823@yahoo.com molecules on expansion of cord blood HSCs.
Abbas Basiri
basiri@unrc.ir Keywords: small molecules, ex vivo expansion, hematopoietic stem cell, expression, mesenchymal stromal cells
Specialty section:
This article was submitted to INTRODUCTION
Stem Cell Research,
a section of the journal Hematopoietic stem cells (HSCs) are a group of cells being produced during embryogenesis
Frontiers in Cell and Developmental to preserve the blood system. Unlike to progenitor cells, these cells have both self-renewal
Biology and multipotency. Therefore, HSCs have important applications in the hematopoietic stem cell
Received: 03 January 2021 transplantations (HSCT) and regenerative medicine (Tajer et al., 2019). However, HSCs constitute a
Accepted: 22 March 2021 minor population of bone marrow cells even less than 0.01% of these cells (Walasek et al., 2012). The
Published: 09 April 2021
fast accessibility and lower need for immune-matching have potentiated the umbilical cord blood
Citation: as an important source of HSCs for transplantation (Chou et al., 2010). Considering the restricted
Ghafouri-Fard S, Niazi V, Taheri M quantities of HSCs in the umbilical cord blood and inadequate mobilization of bone marrow stem
and Basiri A (2021) Effect of Small
cells (Daniel et al., 2016), ex vivo expansion of these HSCs represents an applicable method for
Molecule on ex vivo Expansion
of Cord Blood Hematopoietic Stem
obtaining substantial quantities of HSCs. Several strategies have been used to expand these cells
Cells: A Concise Review. among them is addition of several cytokines to the culture media. Yet, this method did not lead
Front. Cell Dev. Biol. 9:649115. to adequate and long lasting expansions (Zhang and Lodish, 2008). Lack of sustained self-renewal
doi: 10.3389/fcell.2021.649115 and induction of differentiation in HSCs obtained from these protocols (Seita and Weissman, 2010)
Frontiers in Cell and Developmental Biology | www.frontiersin.org 1 April 2021 | Volume 9 | Article 649115Ghafouri-Fard et al. Small Molecule Hematopoietic Stem Cells
have restricted the application of these methods. However, over-expression was accompanied by global hypomethylation
more recent studies have attained promising results using (Saraf et al., 2015). Milhem et al. (2004) have treated human
hematopoietic expansion medium, comprising cytokines and bone marrow CD34+ cells with a cytokine cocktail, 5azaD,
nutritional complements (Zhang et al., 2019). Other modalities and TSA. They observed remarkable expansion of a group of
for ex vivo expansion of HSCs include co-culture with stromal these cells. Notably, 5azaD- and TSA-pre-exposed cells but not
cells (McNiece et al., 2004), forced over-expression of specific those treated with cytokines alone preserved the capacity to
genes (Walasek et al., 2012), and using recombinant proteins repopulate NOD mice (Milhem et al., 2004). After a period
for modulation of developmental pathways (Krosl et al., 2003). of cytokine priming, VPA-exposed CD34+ cells have produced
Besides, lentivirus vectors have been used to deliver a number of CD34+CD90+ multipotent cells. Co-culture of CD34+ cells
genes to enhance engraftment of short term repopulating HSCs with combination of cytokines and VPA has resulted in a more
(Abraham et al., 2016). Numerous small-sized chemical agents remarkable expansion of these cells. Such exposure has led
have also been used for such purpose (De Lima et al., 2008; to enhancement of aldehyde dehydrogenase activity, and over-
Nishino et al., 2009; Peled et al., 2012). In the current manuscript, expression of a number of cell surface proteins namely CD90,
we provide a concise summary of the effects of diverse small CD117, CD49f, and CD184. Treatment of CD34+ cells with VPA
molecules on expansion of cord blood HSCs. has led to production of higher quantities of repopulating cells in
the immune deficient mice (Chaurasia et al., 2014).
STEMREGENIN-1 (SR-1)
NICOTINAMIDE
StemRegenin–1 has been shown to enhance expansion of
CD34+ hematopoietic progenitors through antagonizing aryl As a suppressor of SIRT1, Nicotinamide has been shown to
hydrocarbon receptor (Boitano et al., 2010). Co-culture of HSCs preclude differentiation and enhance expansion of hematopoietic
with SR-1 and several other factors such as stem cell factor stem and progenitor cells (HSPCs) (Peled et al., 2012). Horwitz
(SCF), FLT-3L, TPO, and IL-6 has resulted to expansion of larger et al. (2014) have used an ex vivo–expanded cell product
quantities of CD34+ cells (Wagner et al., 2016). In a clinical trial that contains nicotinamide (NiCord) in a phase I clinical
conducted by Wagner et al. (2016) SR–1 has resulted in a 330- trial. They reported total or fractional neutrophil and T cell
fold expansion of CD34+ cells resulting in fast engraftment of engraftment in most of patients. Moreover, they reported stability
neutrophils and platelets in all of assessed patients. According of NiCord engraftment in all individual during the follow-up
to remarkable effect of this substance on HSCs expansion, non- period. Median neutrophil recovery was faster in individuals
existence of graft failure and high hematopoietic recovery, SR-1 transplanted with NiCord (Horwitz et al., 2014).
has been suggested as a solitary agent for HSCT for defeating
the major problem of umbilical cord blood transplantation
(Wagner et al., 2016).
SEROTONIN
The neurotransmitter serotonin has various extraneuronal roles.
This substance has been shown to induce megakaryocytopoiesis
EPIGENETIC MODIFIERS through 5−HT2 receptors (Yang et al., 1996). In addition,
serotonin promotes expansion of CD34+ cells to early
Mahmud et al. (2014) have assessed expansion of HSCs when stem/progenitors and multilineage committed progenitors.
exposed to histone deacetylase (HDAC) inhibitors valproic acid The effects of this agents on repopulating cells in the expansion
(VPA) and trichostatin A (TSA). These cells were exposed to these culture has also been verified in SCID mice suggesting the impact
agents alone or along with 5-aza-20 -deoxycytidine (5azaD). Their of serotonin in the development of HSCs and modulation of
experiment showed the superior effects of VPA on expansion bone marrow niche (Yang et al., 2007).
of CD34+CD90+ cells and progenitor cells. In vivo studies Table 1 summarizes the results of studies which assessed the
verified the impacts of VPA on prevention of HSC defects. effects of small molecules on ex vivo expansion of cord blood
Besides, combination of 5azaD and TSA resulted in expansion of hematopoietic stem cells.
HSCs that preserve their features through serial transplantation.
Expression analysis revealed differential expression of genes
participating in the expansion and maintenance of HSCs in DISCUSSION
5azaD/TSA- and VPA-treated cells, respectively. Overexpression
of quiescence genes by histone acetylation has been suggested Ex vivo expansion of HSCs obtained from umbilical cord blood
as the underlying mechanism of these observations (Mahmud has become a major research field because of its application
et al., 2014). Saraf et al. (2015) have assess the effects of in the treatment of several hematological disorders (Flores-
sequential treatment of CD34+ mobilized human peripheral Guzmán et al., 2013). This process is expected to mimic the
blood (MPB) with 5azaD and TSA in accompany with cytokines. normal bone marrow niche which is consisted of stromal
They observed significant expansion of CD34+CD90+ cells cells and their produced materials. Thus, nearly all expansion
in 5azaD/TSA-treated cells. They also detected over-expression protocols contain early- and late-acting cytokines to enhance
of genes participating in self-renewal in these cells. Such growth of hematopoietic cells. Moreover, various kinds of
Frontiers in Cell and Developmental Biology | www.frontiersin.org 2 April 2021 | Volume 9 | Article 649115Ghafouri-Fard et al. Small Molecule Hematopoietic Stem Cells
TABLE 1 | Effects of small molecules on ex vivo expansion of cord blood hematopoietic stem cells.
Molecule Input cells Media type and Expansion Result Possible mechanism Reference
cytokines period
SR1 HSC CD34+ – 15 days 330-fold increase in HSC Inhibition of AhR and differentiation of HSC Wagner et al., 2016
CD34+
5azaD/TSA HSC CD34+ Serum-containing media, 9 days 10.7-fold increase in HSC Hypomethylation of self-renewal genes in HSC Mahmud et al., 2014
SCF, Flt3L, IL-3, TPO CD34+ and activation of them
DEAB HSC CD34+ IMDM, 10% FBS, SCF, 7 days – Inhibition of aldehyde dehydrogenase (ALDH) Chute et al., 2006
TPO, Flt3L and downregulation of retinoic acid signaling
pathway and overexpression of cEBP
P18IN003 and HSC CD34+ IMDM/ 10% FBS, SCF, 3 days – Inhibition of sp18INK4C and increase in Srikanth et al., 2015
P18IN011 TPO, Flt3L self-renewal of HSC
TEPA HSC CD34+ α-MEM, 10% FCS, SCF, 21 days – Increase in the activity of cytochrome c Prus and Fibach, 2007
TPO, Flt3L, IL3, IL6 oxidase
Nicotinamide HSC CD34+ – – 104-fold increase in HSC Inhibition of SIRT1 deacetylase and Horwitz et al., 2014
CD34+ differentiation of HSC
Notch ligand HSC CD34+ – 16 days 164-fold increase in HSC Induction of self-renewal by activation of Dahlberg et al., 2011
CD34+ Notch receptor
MSC co-culture Unselected – 11 days 30.1-fold increase in HSC Increase in SDF-1 which diminishes Robinson et al., 2006
CD34+ differentiation of HSC
Nicotinamide HSC CD34+ – 21 days 72-fold increase in HSC Inhibition of SIRT1 deacetylase and Horwitz et al., 2014
CD34+ differentiation of HSC
Fucosylation Unselected – 30 min NA Increase in the homing ability of HSC Popat et al., 2015
5azaD/TSA HSC CD34+ Serum-containing media, 9 days 3.6-fold increase in HSC Hypomethylation of self-renewal genes in HSC Saraf et al., 2015
SCF, Flt3L, IL-3, TPO CD34+ and activation of them
5azaD/TSA HSC CD34+ Serum-containing media, 9 days 2.5-fold increase in HSC Hypomethylation of self-renewal genes in HSC Milhem et al., 2004
SCF, Flt3L, IL-3, TPO CD34+ and activation of them
VPA HSC CD34+ Serum-containing media, 9 days 64.6-fold increase in HSC Overexpression of quiescence genes by Mahmud et al., 2014
SCF, Flt3L, IL-3, TPO CD34+ histone acetylation
SB203580 HSC CD133+ SCF, TPO, Flt3L 7 days – Decline in senescence of HSC, Zou et al., 2012
overexpression of CXCR4
OAC1 HSC CD34+ RPMI, 10% FBS, SCF, 4 days – Overexpression of HOXB4 gene, Induction of Huang et al., 2016
TPO, Flt3L OCT4 expression
dm-PGE2 HSC CD34+ IMDM/ 2% FCS 3, 6, or 9 h – Increase in homing, survival, and proliferation Simsek et al., 2010
and CD133+ of HSCs
VPA HSC CD34+ Serum-free media, 16 h 7 days 20-fold increase in HSC Overexpression of pluripotency genes Chaurasia et al., 2014
priming with SCF, Flt3L, CD34+
IL-3, TPO
VPA HSC CD34+ Serum-containing media, 7 days 89-fold increase in HSC Overexpression of pluripotency genes Chaurasia et al., 2014
SCF, Flt3L, IL-3, TPO CD34+
UM171 HSC CD34+ Fed-batch culture system 7 days 60–75-fold increase in HSC Suppression of erythroid and megakaryocytic Fares et al., 2014
CD34+ differentiation
Notch ligand HSC CD34+ Serum-free media, SCF, 17–21 days 222-fold increase in HSC Stimulation of Notch signaling and induction of Delaney et al., 2010
Flt3L, IL-3, IL-6, TPO CD34+ self-renewal
SR1 HSC CD34+ Serum-free media, SCF, 21 days 270-fold increase in HSC Inhibition of AhR and differentiation of HSC Boitano et al., 2010
TPO, IL-6, Flt3L CD34+
5azaD, TSA HSC CD34+ Serum-containing Media, 9 days 5-fold increase in HSC Unclear Araki et al., 2006
SCF, TPO, FL, IL-3 CD34+
UNC0638 HSC CD34+ Serum-free media, SCF, 14 days 60-fold increase in HSC Unclear Chen et al., 2012
TPO, Flt3L, IL-6 CD34+
NR-101 HSC CD34+ StemSpan/SCF, TPO, Flt3L 7 days – Activation of TPO receptor (c-MPL), activation Nishino et al., 2009
of c-MPL, STAT5 Pathways, overexpression of
VEGF
zVADfmk HSC CD34+ StemPro/SCF, TPO, Flt3L, 10 days – Pan caspase inhibitor, overexpression of Bcl-2 Kale and Limaye, 2010
IL6 and downregulation of Caspase-3, Activation
of notch1
Nicotinamide HSC CD34+ α-MEM/10% FBS, SCF, 21 days – Downregulation of p53 gene Peled et al., 2012
TPO, Flt3L, IL6
Resveratrol HSC CD34+ StemSpan/SCF, TPO, 9 days – Overexpansion of CD34+/CD133+ and cell Heinz et al., 2015
Flt3L, IL6 cycle associated genes, downregulation of
cyclooxygenase 1 (COX-1)
Copper chelator HSC CD133+ – 21 days 6-fold increase in HSC Unclear De Lima et al., 2008
(TEPA) CD133+
dmPGE2 Unselected – 2h NA Increasing Wnt signaling Hagedorn et al., 2014
Garcinol HSC CD34+ StemSpan/SCF, TPO, Flt3L 7 days – Inhibition of HATs Nishino et al., 2011
Serotonin (5-HT) HSC CD34+ QBSF-60/SCF, TPO, Flt3L, 8 days – Activation of 5-HT receptor, Down-regulation Yang et al., 2007
IL6 of Caspase-3
TEPA, Tetraethylenepentamine; 5azaD/TSA, 5aza-20 - Deoxycytidine/Trichostatin A; MSC, Mesenchymal Stromal Cells; VPA, Valproic Acid; FCS, fetal calf serum; dmPGE2,
Dimethyl-Prostaglandin E2; Notch, Notch ligand; HSC, Hematopoietic Stem Cell; SR1, StemRegenin 1; TSA, trichostatin A; AhR, aryl hydrocarbon receptor; SCF,
stem cell factor; SIRT1, Silent Mating Type Information Regulation 2 Homolog 1; IL-6, interleukin 6; TPO, thrombopoietin; SDF1, Stromal Derived Factor-1; DEAB,
diethylaminobenzaldehyde; IMDM, Iscove’s Modified Dulbecco’s Medium; FBS, fetal bovine serum; RPMI, Roswell Park Memorial Institute; VEGF, Vascular endothelial
growth factor; α-MEM, Minimum Essential Medium Eagle – alpha modification; HAT, histone acetyltransferases.
Frontiers in Cell and Developmental Biology | www.frontiersin.org 3 April 2021 | Volume 9 | Article 649115Ghafouri-Fard et al. Small Molecule Hematopoietic Stem Cells
stromal cells such as primary MSCs can improve the efficiency As a rule, strong expansion of HSPCs with sustaining
of expansion. Several promising results have been obtained engraftment requires the synergistic functions of several
from these experiments leading to design of nearly optimal enhancers of HSC expansion (Pineault and Abu-Khader, 2015).
protocols. Different combinations of cytokines can promote This fact has been reflected in better therapeutic results and
commitment of hematopoietic progenitor cells (HPCs) from enhanced engraftment in clinical trials that used the synergistic
certain lineages, i.e., erythroid or myeloid ones. Thus, selection effects of cytokines and HSPC expansion enhancers (Pineault
of appropriate cytokines is important in reaching the optimal and Abu-Khader, 2015). Therefore, application of combination
yield. The possibility of differentiation of HSCs to non- of small molecules and cytokines is a suggested strategy for
hematopoietic cells, including neural, cardiac and myogenic cells expansion of HSCs and inhibition of their differentiation (Wang
have expanded their applications in the treatment of human et al., 2017). The underlying mechanism of such observations
disorders (Flores-Guzmán et al., 2013). Moreover, expanded HSC might be the effect of small molecules in modulation of regulatory
have been used in clinical trials to inhibit Graft-versus-host roles of cytokines on signaling pathways. The results of in vitro
disease (GVHD) following HSCT. Examples of these clinical studies have been verified in a number of experiments using
trials are those conducted by Nohla Therapeutics1 and exciting irradiated NOD/SCID mice (Eldjerou et al., 2010; Budak-
trials by ExCellThera2 . The latter has received Food and Drug Alpdogan et al., 2012). These types of studies would help in
Administration orphan drug designation for ECT-001. ECT-001 the evaluation of cell viability and engraftment capacity in vivo.
comprises a small molecule, UM171, and an adjusted culture Functional consequences of HSC expansion strategies in the
method for expansion of HSC. The primary results of this trial clinical settings have been assessed through different methods.
have been promising in reduction of risk of GVHD3 . The time to neutrophil or platelet engraftments, enhancement
Small molecules represent novel modalities for ex vivo of patients’ survival, occurrence of GVHD, infectious conditions,
expansion of cord blood HSCs. These molecules have also been or relapse rates are among parameters which have been
used for the purpose of expansion of HSPCs with primitive recognized as clinically relevant parameters in this regard
phenotype. Screening of a patented library containing tens of (Kiernan et al., 2017). The most important parameter seems
small molecules has shown the necessity of including SCF, to be patients’ survival. It is worth mentioning that in spite
thrombopoietin and Fms-related tyrosine kinase 3 ligand in the of important developments in ex vivo expansion of HSCs, the
cocktail for enhancing expansion of these cells in addition to C7. capability to follow HSC fate decisions within the bone marrow
Moreover, adding insulin like growth factor binding protein 2 to microenvironment is inadequate (Papa et al., 2020).
the cocktail slightly increased expansion (Bari et al., 2016). Another application of small-molecules-induced expansion of
The efficacy of these molecules is different in various aspects HSCs is that transcriptome analysis of these cells can facilitate
of HSCT. For instance, SR-1, Notch-ligand and nicotinamide- identification of the crucial pathways and molecules of HSC
based methods has been associated with fast neutrophil recovery self-renewal, thus providing novel target molecules for in vitro
(Delaney et al., 2010; Horwitz et al., 2014; Wagner et al., expansion of these cells (Zhang et al., 2020).
2016). Epigenetic modifiers such as VPA are among the mostly Taken together, several molecules such as SR-1, NR-101, the
assessed agents in ex vivo expansion studies. Such agents caspases inhibitor zVADfmk, the calpain inhibitor zLLYfmk,
increase expression of multipotency and self-renewal genes Nicotinamide, Resveratrol, the Copper chelator TEPA, dmPGE2,
mainly through global modifications in epigenetic marks. Garcinol, and serotonin have been used in combination of
Small molecules exert their effects through different routes cytokines to expand HSCs ex vivo. The most promising results
including induction of expression of pluripotency genes, have been obtained from cocktails that influence multipotency
modulation of methylation pattern of self-renewal genes and and self-renewal features from different pathways. These
regulation of retinoic acid, Wnt, and Notch signaling pathways. cocktails include several growth factors, molecules that regulate
Inhibition of apoptosis is another putative mechanism of signaling pathways and vectors that manipulate expression of
participation of small molecules in ex vivo expansion of HSCs, genes. Putative target genes in this regard are HOX genes and
as the caspase and calpain inhibitors have been proved to be cell cycle associated genes as well as those participating in Wnt,
effective in this regard (Kale and Limaye, 2010). Such strategy Notch, and Shh pathways. Each constituent of these cocktails
has been proved to be effective in enhancement of efficacy of might affect some aspects of multipotency and self-renewal,
transplantation in animal models, since temporary up-regulation constructing a synergic system to yield the best results. These
of prosurvival BCL-XL as considerably promoted survival and elements should enhance self-renewal, suspend differentiation,
engraftment of HSCs and progenitor cells (Kollek et al., 2017). promote homing, and suppress apoptosis of HSCs (Zhang and
In some cases, the exact molecular mechanism of their impacts Gao, 2016). Precise assessment of the expansion aptitude of cell
on HSCs is not clear. Identification of such mechanisms would populations through quantification of the generated cells using
facilitate design of optimal protocols and avoidance of possible immunophenotypic methods or functional assessments is an
side-effects of associated therapies. important requirement of comparison studies for identification
of best ex vivo expansion methods. Aptitude of expansion and
1
https://clinicaltrials.gov/ct2/show/NCT01690520
preservation of multipotent HPCs can be precisely assessed by
2
https://www.clinicaltrials.gov/ct2/show/NCT04103879 the numbers of lineage specific colony-forming units and colony-
3
https://www.insidertracking.com/excellthera-receives-fda-orphan-drug- forming unit mix and Human Long-Term Culture-Initiating Cell
designation-for-ect-001-for-the-prevention-of-graft-versus-host-disease (LTC-IC) activity (Ali et al., 2014). Concomitant conduction of
Frontiers in Cell and Developmental Biology | www.frontiersin.org 4 April 2021 | Volume 9 | Article 649115Ghafouri-Fard et al. Small Molecule Hematopoietic Stem Cells
these methods is crucial for identification the expanded lineage in proteins (Deutsch et al., 2010; Celebi et al., 2011) or Sonic
each experimental condition. hedgehog (Bhardwaj et al., 2001) and induction of expression
As different methods have been used for such quantifications, of genes which promote self-renewal (Watts et al., 2010;
it is not easy to precisely compare the results of different Aguila et al., 2011).
studies. Most mentioned studies has used CD34+ HSCs Finally, documentation of expansion of actual HSCs rather
as input cell population. Yet, a number of studies lave than HPCs is an important prerequisite of clinical applications
used unselected cell populations which complicate the although both cell population have their own clinical uses. Such
comparison studies due to different extents of enrichment for important step has been missed in some early studies in this field.
primitive cells.
Considering the complexity of natural HSC milieu and
existence of a wide range of cells and their produced AUTHOR CONTRIBUTIONS
molecules in addition to secreted cytokines, application of
multifaceted strategies seems to yield more promising results. MT and SG-F wrote the draft of the manuscript and revised it.
Examples of such strategies are co-culture of stromal cells AB and VN designed the tables and collected the data. All authors
with other niche components including extracellular matrix approved the submitted version.
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Superior ex vivo cord blood expansion following co-culture with bone marrow- absence of any commercial or financial relationships that could be construed as a
derived mesenchymal stem cells. Bone Marrow Transp. 37, 359–366. doi: 10. potential conflict of interest.
1038/sj.bmt.1705258
Saraf, S., Araki, H., Petro, B., Park, Y., Taioli, S., Yoshinaga, K. G., et al. (2015). Copyright © 2021 Ghafouri-Fard, Niazi, Taheri and Basiri. This is an open-access
Ex vivo expansion of human mobilized peripheral blood stem cells using article distributed under the terms of the Creative Commons Attribution License
epigenetic modifiers. Transfusion 55, 864–874. doi: 10.1111/trf.12904 (CC BY). The use, distribution or reproduction in other forums is permitted, provided
Seita, J., and Weissman, I. L. (2010). Hematopoietic stem cell: self-renewal versus the original author(s) and the copyright owner(s) are credited and that the original
differentiation. Wiley Interdiscip. Rev. Syst. Biol. Med. 2, 640–653. doi: 10.1002/ publication in this journal is cited, in accordance with accepted academic practice. No
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